泌尿道致病性大肠埃希菌Ⅰ型PapG黏附素基因的扩增与分析

Amplification and Identification of ⅠType of papG Gene from Uropathogenic Escherichia coli

目的获得Ⅰ型黏附素代表菌株———泌尿道致病性大肠埃希菌(UPEC)J96 PapG全基因扩增片段并进行克隆子分析。方法设计合成一对可扩增Ⅰ型papG全长基因的PCR引物,采用长片段PCR扩增法获得该基因的扩增片段,以限制性内切酶酶解后克隆pUC18质粒载体。结果限制性酶谱、PCR和序列分析表明,所获得重组质粒pJG29中已带有Ⅰ型papG全长基因片段的克隆。结论所设计合成的引物和建立的PCR扩增法可用于大量获取并分析UPECⅠ型黏附素基因。

Objective To amplificated and cloned the 1 type of papG adhesin gene of uropathogenic Esherichia coli（UPEC） strain J96 was subjected to PCR amplification and molecular cloning, Methods The polymerase chain reaction was used to obtain the DNA fragment of papG gene from UPEC J96. Restriction endonuclease and sequence analysis was used to identify the recombinant plasmid. A pair of oligonucleotide primers were designed according to the DNA sequence of the papG of UPEC J96, and the papG gene was amplified and cloned into plasmid pUC18. Recombinant plasmids were identified by restriction endonuclease and PCR amplification. Results The recombinant plasmid carrying about 1 100 bp fragment of 1 type papG of UPEC J96 was designated pJG29. The sequence analysis revealed that the DNA sequence of papG gene in pJG29 was 1080 bp and correct in comparison with that showed in ＂Gene Bank＂. Conclusion The 1 type of papG gene is obtained by PCR amplification and cloned. The results could provide a useful model for the study of PapG adhesin variants and their pathogenesis in Escherichia coli.